Detergent compositions containing bacillus agaradhaerens mannanase and methods of use thereof
A mannanase, detergent technology, applied in detergent compositions, detergent compounding agents, biochemical equipment and methods, etc., can solve the problems of unsuitability for preparations and washing conditions, etc.
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[0246] The following examples are provided to demonstrate and illustrate certain preferred embodiments and aspects of the invention and should not be construed as limiting.
[0247] In the following experimental disclosures, the following abbreviations are used: M (mole / liter), mM (millimol / liter), μM (micromoles / liter), nM (nanomoles / liter), mol (mole), mmol (mmol ), μmol (micromole), nmol (nanomole), g and gm (gram), mg (milligram), μg (microgram), pg (picogram), L (liter), ml and mL (milliliter), μl and μL (microliter), cm (centimeter), mm (millimeter), μm (micrometer), nm (nanometer), U (unit), MW (molecular weight), s (second), min (minute), h (hour ), ℃ (Celsius), QS (sufficient), ND (not performed), rpm (rev / min), H 2 O (water), dH 2 O (deionized water), HCl (hydrochloric acid), aa (amino acid), bp (base pair), kb (kilobase pair), kD (kilodalton), MgCl 2 (magnesium chloride), NaCl (sodium chloride), Ca (calcium), Mg (magnesium), HEPES (4-(2-hydroxyethyl)-1-piperazine...
example 1
[0249] Cloning of Glycosyl Hydrolase Bag Man1 from Bacillus agaricus
[0250] Bacillus agaricus was selected as a potential source of a variety of glycosyl hydrolases and other enzymes that can be used in industrial applications. Genomic DNA for sequencing was obtained as follows: first, a strain of Bacillus agaricus was placed on a GAM agar plate at 30°C (Jones et al., IJSEM, 55:1711-1714, 2005 (Jones et al., "International Journal of Systematic and Evolutionary Microbiology, Vol. 55, pp. 1711-1714, 2005)) Incubation for 24 hours. Cellular material was scraped from the plates and used to prepare genomic DNA with the aid of the ZF Fungal / Bacterial DNA Miniprep Kit from Zymo (Cat# D6005). The genomic DNA was used for sequencing and used to amplify the Bag Man1 gene for expression cloning. use Sequencing by synthesis (SBS) technology (www.baseclear.com / sequencing / illumina-sequencing) Whole-genome sequencing of Bacillus agaricus viscosus. Genome sequencing and sequence dat...
example 2
[0256] Expression of Glycosyl Hydrolase (Bag Man1) from Bacillus agaricus
[0257] The Bag Man1 gene was amplified by PCR from Bacillus agaricus gluei genomic DNA using the following primers: Primer 1 (BssHII) 5'-TGAGCGCGCA GGCTGCTGGA AAAATGACAA ACTATGAATC AGAGGT-3' (SEQ ID NO: 8), and Primer 2 (BamHI) 5 '-TGTGGATCCT TACTCTAACG GTACGTCTTC CTTAT-3' (SEQ ID NO: 9). The amplified Bag Man1 gene was cloned into the expression vector p2JM by BssHII / BamHI double enzyme digestion and ligation. The Bacillus subtilis expression vector p2JM103BBI (Vogtentanz, Protein Expr Purif, 55:40-52, 2007) was digested with restriction enzymes BssHII and BamHI (Vogtentanz, "Protein Expression and Purification", Vol. 55, pp. 40-52, 2007)). A DNA fragment free of the BCE103-BBI fusion gene sequence was isolated and used as the expression backbone. This DNA fragment was ligated with the PCR amplified gene encoding the BagManl mature protein, which resulted in the addition of three codons between t...
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